His Tag Aptamer Assay

6H5 aptamer: 5’-GGCTTCAGGTTGGTCTGGTTGGGTTTGGCTCCTGTGTACG-3’

HMBA (His-tag molecular beacon aptamer ): 5’-FAM-CAGGTTGGTCTGGTTGGGTTTGGCTCCTGTGTACG-(CH2CH2O)₃₆-CAACCTG-Dabcyl-3’

PEG linker: (CH2CH2O)₃₆

HMBA contains a major part of 6H5 with the deletion of 5 nucleotide residues from the 5-prime end, and addition of a partial cDNA sequence containing 7 nucleotide residues to the 3-prime end and PEG36 as a linker.

FAM was chosen as the fluorophore, and Dabcyl was used as the quencher.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3544214/

FAM was chosen as the fluorophore, and Dabcyl was used as the quencher. anti His tag aptamer : 6H7, 5′-GCT ATG GGT GGT CTG GTT GGG ATT GGC CCC GGG AGC TGG C-3′, and 6H5, 5′-GGC TTC AGG TTG GTC TGG TTG GGT TTG GCT CCT GTG TAC G-3′) and

anti-streptavidin aptamer : (miniStrep, 5′-TCT GTG AGA CGA CGC ACC GGT CGC AGG TTT TGT CTC ACA G-′3)

Aptamer Immobilization. Aptamers were spotted on PEI- modified glass slides or on UltraGAPS Coated Slides in the same

buffer they were activated in (SBB or PBS-6H7, respectively). Dilution rows of the activated aptamers were spotted in 10 replicates of every dilution using a 427 arrayer (Affymetrix, Santa Clara, CA) equipped with 125-μm solid pins. Spotting was performed by placing five hits per dot. After spotting, the slides were incubated at room temperature overnight in the dark. Capping of Amino Modifications. The printed microarrays were incubated in a fresh solution of 0.275 g of succinic acid in